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Complex amplification system applied to 23 human STR sites, kit and application of kit

A complex amplification system, D21S11 technology, applied in the biological field, achieves the effects of high accuracy, high polymorphism, and high stability

Active Publication Date: 2018-01-09
BGI FORENSIC TECH (SHENZHEN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the above 13 CODIS sites are not completely suitable for Chinese ethnic groups

Method used

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  • Complex amplification system applied to 23 human STR sites, kit and application of kit
  • Complex amplification system applied to 23 human STR sites, kit and application of kit
  • Complex amplification system applied to 23 human STR sites, kit and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Utilize the fluorescence-labeled composite amplification kit of 23 STR sites of the present invention to conduct blood samples from a doublet (father-son) and triplet (parent-daughter) family (the sample has been identified and confirmed by the Haiershi HGT 21G kit) to test.

[0076] The family blood samples were donated by volunteers, and the DNA was extracted by the chelex-100 method (for specific methods, refer to "Forensic DNA Protocol", Humana Press, 1998). Take 3 μL of blood with anticoagulants in a 1.5ml centrifuge tube, shake and mix the chelex Solution, to make chelex fully suspended, add 200μL 5% chelex-100 to each tube, shake the sample, after incubating on a constant temperature metal bath at 56°C for 2 hours, take out the sample and shake for 2 minutes, boil for 8-10 minutes, centrifuge at 13000rpm for 3 minutes, be careful Aspirate about 150 μL of supernatant and transfer to a new 1.5ml centrifuge tube. Using the kit of the present invention and Hirsch's ...

Embodiment 2

[0101] Multiple amplification and typing were performed on blood sample 1, where blood sample 1 was donated by volunteers, and the adjacent alleles of D1S1656 in blood sample 1 could not be correctly typed using the existing HGT 21G kit.

[0102] The DNA was extracted by the chelex-100 method (refer to "Forensic DNA Protocol" for specific methods, HumanaPress, 1998), and the sample was amplified and detected simultaneously using the kit of the present invention and the HGT 21G kit of Haiers Gene Technology Co., Ltd. The amplification reaction was carried out on the G1000 thermal cycler, the electrophoresis and detection were carried out on the ABI 3500 genetic analyzer, and the data analysis was carried out by Gene IDx software. The reagents and materials used, such as formamide, internal standard, etc., are conventional materials commonly used by those skilled in the art.

[0103] 2.1 DNA extraction by Chelex-100 method

[0104] 2.2 Polymerase chain reaction (PCR) amplific...

Embodiment 3

[0114] Multiplex amplification and typing were performed on the three loci of blood sample 2 (including sample A, sample B, and sample C). Blood sample 2 was donated by volunteers, and each blood sample 2 contained common microvariant alleles, which could not be correctly typed using the existing MR23sp kit from Yuewei Gene Company.

[0115] DNA adopts the chelex-100 method to extract (specific method with reference to " Forensic DNA Protocol ", HumanaPress, 1998), utilizes kit of the present invention, the kit MR23sp of Yuewei Gene Company to amplify above-mentioned sample 2 simultaneously, amplification reaction is heated in G1000 It was carried out on a cycler, electrophoresis and detection were carried out on an ABI 3500 genetic analyzer, and data analysis was performed using GeneMapperIDx software. The reagents and materials used, such as formamide, internal standard, etc., are conventional materials commonly used by those skilled in the art.

[0116] 3.1 DNA extraction ...

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Abstract

The invention discloses a complex amplification system applied to 23 human STR sites, a kit and an application of the kit. 23 loca, namely D10S1248, D5S818, D21S11, D1S1656, AMEL, D3S1358, D13S317, D7S820, D16S539, D19S433, D22S1045, D2S441, D12S391, D2S1338, D18S1364, SE33, Penta-D, D11S2368, D13S325, D6S1043, Penta-E, Y indel and Y GATA H4, can be simultaneously amplified; the complex amplification system can satisfy genetic polymorphism of the Chinese nation, and the complex amplification system is compatible with core sites of foreign countries; the complex amplification system has the characteristics of being high in polymorphism, high in stability, high in information amount and high in accuracy; and the complex amplification system can be well applied to individual recognition, paternity identification, population genetic analysis and / or construction of human DNA database.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the detection technology of polymorphic genetic marker genes in the human genome, in particular to a system and detection system for simultaneously amplifying 23 loci in a composite amplification system by polymerase chain reaction Kits and their uses. Background technique [0002] Short tandem repeats (short tandem repeats, STRs) are a class of molecular genetic markers widely present in eukaryotic genomes. It has a large amount of information, accounting for about 10% of the entire genome. Generally, the core repeating unit is composed of 2 to 6 bases, and the repeating number of the core unit varies among different individuals, which has high polymorphism and follows the Mendelian law of inheritance. In addition, STR fragments are small, about 70-500 bp, easy to amplify, and suitable for some trace or degraded samples. Therefore, STR-based amplification detection technology is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
Inventor 李生斌伏东科熊文娟李栋曾柳眉李波
Owner BGI FORENSIC TECH (SHENZHEN) CO LTD
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